Application
Western blot
Sample
Dog Cell lysate - whole cell (Canine cell lines, tissues and recombinant canine)
Gel Running Conditions
Reduced Denaturing (7.5%)
Loading amount
50 µg
Specification
Canine cell lines, tissues and recombinant canine
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Other product details
Incubation time
20 hour(s) and 0 minute(s) · Temperature: 4°C · Diluent: 5% BSA in TBST
Dilution
1/500
Secondary antibody
Dilution
1/2000
Name
Non-Abcam antibody was used: Dako Polyclonal goat anti rabbit HRP
Host species: Goat
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Host species: Goat
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Detection
Detection method
ECL
Exposure
28 minute(s) and 0 second(s)
Bands
Specific: 18 kDa Non-specific: approx 105 kDa
Positive control
DO64 cells, C2 cells and testis have all been reported to express stem cell factor protein.
Recombinant canine stem cell factor.
All cell lines and tissues used have been demonstrated to express variable amounts of both membrane bound (approx 46kDa) and soluble (approx 50 kDa) SCF using R&D systems mAb2268. Cell lines expressed varying degrees of soluble and membrane bound mRNA by qPCR.
Additional data
Additional Notes
The predominant band was non specific. Expected bands at 46 and 50kDa were not visible.
D064 bone marrow stromal cell line.
C2 Mast cell tumour line.
Sh1b and CMT-9, 12 ,47, U27, U131 and U229 are all canine mammary epithelial or myoeptihelial cell lines.
T1 - T3 canine testis tissue lysates from separate dogs.
Abcam response
This product is yet to be tested with dog samples, a free product has been offered in exchange of this review as it was tested by end user under our Abtrial offer. We recommend including human cell line lysates as a control lysates.
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.
Abcam user community
Verified customer
提交于 Nov 16 2017